Objective To detect the expression of T helper cells 9 (Th9) in the peripheral blood and the level of interleukin (IL)-9 in the plasma of rheumatoid arthritis (RA) patients,and to analyze the relationship between Th9 and clinical characteristics,and their possible role in the immunological pathogenesis of RA were explored.Methods Thirty-six patients with RA and 22 healthy controls were included.RA patients were divided into high disease activity group (n=22) and moderate disease activity group (n=14).Th9 cells were obtained from RA patients and healthy controls and detected by flow cytometery.The levels of IL-9 in RA patients and healthy controls were detected by ELISA tests.T-test and Spearman's correlation were used for statistical analysis.Results The expression of Th9 and the level of IL-9 of RA patients [(0.99±0.50)％,( 1.06±0.42 ) pg/ml,respectively] were significantly higher than normal controls[ (0.21±0.08)％,(0.69±0.13)pg/ml,respectively,all P＜0.01 ].Meanwhile,the expression of Th9 and the level of IL-9 in high disease activity group [ ( 1.18±0.52 )％,( 1.37±0.47 ) pg/ml ] were higher than those of the moderate disease activity group [ (0.69±0.25)％,(0.94±0.30) pg/ml ].There were positive correlations between Th9 and disease activity score(DAS) 28,erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),the number of tender joints,the number of swollen joints and rheumatoid factor (RF),the level of IL-9 respectively.There were positive correlations between IL-9 and DAS28,ESR,the number of tender joints,and the number of swollen joints.Conclusion High expression of Th9 and level of IL-9 maybe involved in the pat-hogenesis of RA,and is associated with disease activity.These results indicate that the abnormality of Th9 may play an important role in the pathogenesis of RA.

Objective To investgate the expression of Tfh and Th9 in the peripheral blood of rheumatoid arthritis ( RA),and to analyze the relationship between those cells and disease activity,organs of involvement,ect.,and to underly the possible immunological mechananism of RA.Methods Peripheral blood Tfh cells and Th9 cells,define as the CD4+ CXCR5 + ICOS+ and CD3 + CD8+ IL-9+ population,were examined by flow cytometry in 36 patients with RA and 22 case of age-and gender-matched health controls.According to 28-joint Disease Activity Score (DAS28),patients were divied into two groups:high disease activity(n=22),moderate disease activity(n=14).And then to analyzed the correlation among the Tfh,Th9 and clinical data of RA,such as ESR,CRP,RF,the number of tender joints,the number of swollen joints and bone destruction,etc..While to analyzed the correlation between Th9 and Tfh.Results The expression of Tfh (Z=-6.082,P=0.000) and Th9 (0.989±0.498 vs 0.213±0.084,t=13.063,P =0.000 )in PBMCs of RA was significantly higher than normal controls.Meanwhile,the expression of Tfh ( 3.880 ±1.255 vs 2.678±1.022,t=2.990,P=0.005 ) and Th9( 1.181 ±0.523 vs 0.686±0.254,t =4.043,P=0.000) in high activity of group were higher than those moderate.The percentage of Tfh and Th9 in the two groups were all signifieantly higher than controls(P＜0.01 ),There was a positive correlation with Tfh and DAS28 ( r =0.571,P=0.000),ESR ( r =0.375,P =0.029),CRP( r =0.357,P =0.032 ),the number of tender joints ( r=0.598,P =0.000 ),RF ( r =0.421,P =0.023 ) and anti-CCP ( r =0.421,P =0.023 ).There were no relationship with Tfh and course,morning stiffness,the number of swollen joints,bone erosion and the abnormal of EKG.There was a positive correlation with Th9 and DAS28( r=0.461,P=0.005 ),ESR(r=0.347,P=0.042),CRP(r=0.384,P=0.210),the number of tender joints(r=0.341,P=0.042),the number of swollen joints (r =0.347,P=0.038 ) and RF( r =0.379,P =0.025 ) respectively.There were no relationship with Th9 and course,morning stiffness,CCP,the abnormal of EKG,bone erosion.There was a positive correlation with Tfh and Th9 (r=0.727,P =0.000),Conclusion The expression of Tfh and Th9 were increase significantly,which those cells were close relationship with the disease activity and some data,These results indicate that the abnormality of Tfh and Th9 may play an important role in the pathogenesis of RA.

Sepsis is defined as that a variety of pathogenic microorganisms(including bacteria,fungi,viruses and protozoa) invade the blood circulation,produce toxins and cause systemic infection.The early symptoms of neonatal sepsis are atypical.Neonatal sepsis is urgent and progresses quickly,especially in preterm infants,and it is easy to prone to septic shock which is life-threatening.Therefore,early identification,accurate diagnosis,and active intervention for neonatal sepsis and septic shock is essential to reduce mortality and improve prognosis.

Objective To investigate the mastery of clinical students' doctor-patient communi-cation skills and to assess the teaching effectiveness about the doctor-patient communication, and then propose some improvement ideas. Methods We conducted a study in students of clinical medicine of Nanjing Medical University in Grade 2009, using Objective Structured Clinical Examination (OSCE) after internship, which included graduation assessment of doctor-patient communication skills. Then we analyzed the doctor-patient communication skills and related scores of students trained in different clinical medical programs. Results The average score of doctor-patient communication skills of 549 students was (82.72±4.23), of which, the average score of 329 five-year students of clinical medicine was (81.88±4.23) and the average score of seven-year students was (83.96±3.91) in. The average score of seven-year students of clinical medicine was significantly higher than five-year students (P=0.000). Moreover, there was a significant positive correlation between the total score of OSCE and the score of doctor-patient communication skills in both five-year and seven-year students of clinical medicine (five year program students' score correlation coefficient was 0.520, P=0.000;seven year pro-gram students' score correlation coefficient 0.416, P=0.000). Conclusion The teaching effectiveness of doctor-patient communication has proved to be quite effective, and it is definitely of great significance in improving clinical students' doctor-patient communication skills. The score of the assessment of the doctor-patient communication reflects not only the training effectiveness of the communication skills, but also the comprehensive capacity.

To study the effect of oxygen enrichment of room air on the change of hemorheology of soldiers after high altitude hypoxia and exhaustion exercise at 3700 altitude, the oxygen concentration was increased to 24 0%～25 0% in the rooms at 3700m altitude, the change of hemorheology in 10 soldiers after they had 12h rest and sleep in the oxygen enrichment room was examined in no oxygen enrichment resting state and in oxygen enrichment state as well as after exhaustion exercise in no oxygen enrichment room and in oxygen enrichment room.Hematocrit (HCT), blood viscosity (?b), reduction viscosity (?r) and viscometric aggregation index (VAI) were higher after exhaustion exercise in no oxygen enrichment room as compared with those in no oxygen enrichment resting state, the difference was very significant or signifant ( P 0 05),?b, Plasma viscosity (?p), ?r were decreased after exhaustion exercise in no oxygen enrichment room ( P

Objective To explore the efficacy of pushen capsule on blood lipid and atherosclerosis in patients with ischemic stroke.Methods One hundred and twenty patients with ischemic stroke were randomly divided into three groups,treatment group (pushen capsule and rosuvastatin calcium),control group-1 (pushen capsule) and control group-2 (rosuvastatin calcium).There were 40 patients for each group.The course of treatment was 6 months.Data of serum total cholesterol (TC),triacylglycerol (TG),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C),homocysteine (Hcy),high sensitive C reactive protein (hs-CRP),nitric oxide (NO),endothelin-1 (ET-1),the number and size of carotid plaques,and changes of intima-media thickness (IMT) were detected before and after treatment in three groups.The incidences of adverse cerebrovascular events and adverse events were observed in the three groups.And the therapeutic effects for dyslipidemia were evaluated in three groups.Results The treatment indicators were obviously improved after treatment in three groups (P ＜ 0.05).There were no significant differences in treatment indicators before treatment between three groups.The levels of TG and NO increased,the level of LDL-C,the size and thickness of carotid plaques decreased after treatment in control group-2 than those of control group-1.Levels of TC,LDL-C,Hcy,hs-CRP,ET-1,IMT,and the number,size and thickness of carotid plaques were significantly lower in treatment group than those of control group-1 (P ＜ 0.05),and the level of NO was higher in treatment group than that of control group-1 (P ＜ 0.05).Compared with control group-2,levels of TG,LDL-C,hs-CRP and the number,size and thickness of carotid plaques were significantly lower,and levels of HDL-C and NO were significantly increased in treatment group (P ＜ 0.05).There were no significant differences in the incidences of adverse drug reactions and adverse cerebrovascular events between the three groups (P ＞ 0.05).The total effective rate was higher in treatment group than that of control group-1 (P ＜ 0.05).There were no significant differences in total effective rates between treatment group and control group-2,and between control group-2 and control group-1 (P ＞ 0.05).Conclusion Pushen capsule combined with rosuvastatin calcium can obviously improve blood lipid metabolism in patients with ischemic stroke,which shows obvious antiatherosclerosis effect and good safety.

Objective To investigate the effect of salvia mihiorrhiza on spinal cord ischemia reperfusion injury after surgical treatment in patients with cervical canal stenosis. Methods Retrospective analysis of 64 cases had cervical canal stenosis in the last 5 years in our hospital. Sixty-four cases were randomly divided into the salvia mihiorrhiza group(31 cases)and the control group(33 cases). The therapeutic effect was assessed using JOA grade system. Results In the salvia mihiorrhiza group,the JOA average score was 8. 8 ±2. 6 before surgical treatment, after two weeks of surgical treatment it was 13. 7 ± 2. 4. The JOA improvement ratio was (61. 5 ± 2. 9) % . In the control group,the JOA average score was 9. 1 ±2. 2 before surgical treatment,after two weeks of surgical treatment it was 13. 4 ± 2. 3. The JOA improvement ratio was (60. 5 ± 2.2)% .The JOA improvement ratio in the salvia mihiorrhiza group was significantly higher than that in the control group (P < 0. 05) . Conclusions Salvia mihiorrhiza has protective effect on spinal cord ischemia reperfusion injury.

In a previous study, we cloned popW from Ralstonia solanacearum strain ZJ3721, coding PopW, a new harpin protein. The procaryotically expressed PopW can induce resistance to Tobacco mosaic virus (TMV), enhance growth and improve quality of tobacco, when sprayed onto tobacco leaves. Here, we constructed an expression vector pB- popW by cloning popW into the bionary vector pBI121 and transformed it into Agrobacterium tumefaciens strain EHA105 via freeze-thaw method. Tobacco (Nicotiana tobacum cv. Xanthi nc.) transformation was conducted by infection of tobacco leaf discs with recombinant A. tumefaciens. After screening on MS medium containing kanamycin, PCR and RT-PCR analysis, 21 T3 lines were identified as positive transgenic. Genomic intergration and expression of the transferred gene were determined by PCR and RT-PCR. And GUS staining analysis indicated that the protein expressed in transgenic tobacco was bioactive and exhibited different expression levels among lines. Disease bioassays showed that the transgenic tobacco had enhanced resistance to TMV with biocontrol efficiency up to 54.25%. Transgenic tobacco also exhibited enhanced plant growth, the root length of 15 d old seedlings was 1.7 times longer than that of wild type tobacco. 60 d after transplanting to pots, the height, fresh weight and dry weight of transgenic tobacco were 1.4, 1.7, 1.8 times larger than that of wild type tobacco, respectively.
Agrobacterium tumefaciens ; Animals ; Bacterial Outer Membrane Proteins ; genetics ; Disease Resistance ; genetics ; Genetic Vectors ; Phenotype ; Plant Diseases ; prevention & control ; virology ; Plants, Genetically Modified ; genetics ; Tobacco ; genetics ; Tobacco Mosaic Virus ; Transformation, Genetic

ObjectiveTo establish an ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for detecting the bile acid expression in serum and to explore the relationship between the bile acid spectrum and the obesity.Methods After pre-treatment through solid phase extraction,serum samples were chromatographed by gradient elution on an UPLC HSS T3 column,and then mass spectrometric analysis of bile acid was performed by multiple reaction monitoring ( MRM ) mode with negative electrospray ionization (ESI).And its methodology performance, including specificity,linearity,sensitivity,imprecision and recovery,were verified according to the guideline of State Food and Drug Administration (SFDA).Furthermore,serum samples from 10 simple obesity subjects and 10 healthy controls were analyzed with this establishedmethod, and ranksum test was used for statistical analysis.Results This established UPLC-MS/MS method could simultaneously quantify 14 bile acid subgroups in serum sample,its analytical linear range was 10 - 20 000 nmol/L.For different bile acid subgroups,the limit of detection (LOD) was 0.02 - 7.90 nmol/L,limit of quantitation (LOQ) was 0.07 -44.20 nmol/L; and its within-day coefficient of variation (CV) was 0.35％ - 12.41％,between-day CV was 1.11％ - 13.04％ ; the relative recovery ratio of this method was 89.8％ - 114.6％.Some differenceswere observed in chromatogram between obesity and control subjects,and both free bile acid and conjugated bile acid concentrations in obesity subjects [ 0.49 ( 0.45 - 1.90 ),1.44 ( 0.84 - 3.72 ) μmol/L ] were lower than them in control subjects [0.98(0.53 -3.06),1.99(0.67 -2.88) μmol/L],but the difference was not significant ( Z =- 0.958,- 0.801,P ＞ 0.05 ).ConclusionsThis established UPLC-MS/MS method can efficiently differentiate and quantify 14 bile acid subgroups,and is characterized with broad analytical measurement range,good analytical sensitivity and precision.This method can be applied for the free and conjugated bile acid analysis in clinical specimens.

Objective To study the effects of up-regulated gene-4 (URG-4) on colon cancer cellproliferation.MethodsColon cancer cell line with high expression of URG-4 was selected.The recombinant URG-4 siRNA retroviral vector was constructed and packaged by PT67 cell,then retroviral particles which can express URG-4 siRNA in mammal cell and its negative control were obtained.Expressions of URG-4 in MKN45,SW480,LoVo,HCT116,HT29 were detected by RT-PCR and Western blot,respectively.Recombinant virus (interference group),original virus (negative control group) and the same amount of PBS (blank group) were used to transfect LoVo cells respectively.Stably transfected cell lines were screened.The growth condition of cell lines in each group was assayed by MTT.All data were analyzed by the one-way analysis of variance and the t test.Results Sequencing results confirmed the successful construction of retroviral which expressed siRNA,the relative expression levels of URG-4 mRNA in MKN45,SW480,LoVo,HCT116,HT29 were 0.58 +0.02,0.63 ±0.03,0.81 ± 0.01,1.01 ± 0.02,0.91 ± 0.04 and the expression levels of URG-4 protein in the 5 cell lines were 0.73 ±0.02,0.85 ± 0.03,1.42 ± 0.01,0.80 ± 0.30,0.80 ± 0.04,respectively.High expression of URG-4 was observed in the LoVo cells.The expression of URG-4 mRNA in the LoVo cells in the interference group was 0.55 ±0.03,which was significantly lower than 1.15 ±0.02 of the negative control group and 1.15 ±0.01 of the blank group ( t =- 5.179,- 9.285,P ＜ 0.05 ).The inhibition rate of URG-4 mRNA in the interference group was 52.6％.The expression of URG-4 protein in the interference group was 0.82 ± 0.05,which was significantly lower than 1.46 ± 0.07 of the negative control group and 1.54 ± 0.04 of the blank group (t =-4.239,-3.704,P＜0.05).The inhibition rate of URG-4 protein in the interference group was 43.6％.The LoVo cells in each group grew exponentially.Compared with the negative control group,the cell growth of the interference group was inhibited during day 3 to day 6,which had statistical significant difference ( t =- 6.436,-6.045,-6.434,-4.285,P＜0.05).ConclusionInterference of the expression of URG-4 can inhibit the growth of LoVo cells.